USPatent publicationPublished

Aminoquinoline and aminopyridine derivatives and their use as adenosine a3 ligands

Published 23 Sep 2004 · application patented

Assignee: Sanofi

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Inventors: Olivier Finance, Peter Arányi, Endre Mikus, Geza Tímári +10 · Examiner: D. Margaret Seaman · AU 1625 · TC 1600

Application
10/478,721
filed 29 May 2002
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US 20040186133 A1
published 23 Sep 2004
Patent
US 6,969,723
granted 29 Nov 2005
23 Sep 2004
Published
US pre-grant publication
20
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1 independent
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A61K/, A61P11/00
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Olivier Finance
Patented
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granted 29 Nov 2005
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Abstract

Compounds of general formula (I), wherein R 4 and R 5 stand for hydrogen atom or form together an 1,3-butadienyl group, optionally substituted by a methylenedioxy group or one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, hydroxy group or halogen atom; are strong adenosine A 3 receptor ligands preferably antagonists. [structure]

Description

15 parts
›The present invention relates to adenosine A 3…

The present invention relates to adenosine A 3 receptor ligands of the general formula (I), within those preferably antagonists, as well as their salts, solvates and isomers, and the pharmaceutical compositions containing them, to the use of the compounds of the general formula (I), as well as their salts, solvates and isomers, to the preparation of the compounds of the general formula (I) and their salts, solvates and isomers, furthermore to the new intermediates of the general formulae (II) (III) and (IV) and to the preparation thereof.

Adenosine is a well-known component of several endogenous molecules (ATP, NAD + , nucleic acids). Besides, it plays an important regulatory role in many physiological processes. The effect of adenosine on heart function was discovered already in 1929. (Drury and Szentgyörgyi, J Physiol 68:213, 1929). The identification of an increasing number of physiological functions mediated by adenosine and the discovery of new adenosine receptor subtypes give possibilities for therapeutic application of specific ligands (Poulse, S. A. and Quinn, R. J. Bioorganic and Medicinal Chemistry 6:619, 1998).

To date, the receptors for adenosine have been classified into three main classes: A 1 , A 2 and A 3 . The A 1 subtype is partly responsible for inhibiting the adenylate cyclase by coupling to G i membrane protein, partly influences other second messenger systems. The A 2 receptor subtype can be subdivided into two further subtypes—A 2a and A 2b —, which receptors stimulate the adenylate cyclase activity. The sequence of adenosine A 3 receptors have been recently identified from rat testis cDNA library. Later it was proved that it corresponds to a novel, functional adenosine receptor. The activation of the A 3 receptors is connected also with several second-messenger systems: inhibiting of adenylate cyclase, stimulating of phospholipase C and D.

The adenosine receptors are found in several organs and regulate their functions. Both A 1 and A 2a receptors play important roles in the central nervous system and cardiovascular system. In the CNS, the adenosine inhibits the release of synaptic transmitters which effect is mediated by A 1 receptors. In the heart, also the A 1 receptors mediate the negative inotropic, chronotropic and dromotropic effects of adenosine. The adenosine A 2a receptors located relatively in a higher amount in the striatum, display a functional interaction with dopamine receptors in regulating the synaptic transmission. The A 2a adenosine receptors on endothelial and smooth muscle cells are responsible for adenosine-induced vasodilation.

On the basis of mRNA identification, the A 2b adenosine receptors are widely distributed in different tissues. They have been identified almost in every cell type, but its expression is the highest in the intestine and the bladder. This subtype probably also has important regulatory function in the regulation of the vascular tone and plays a role in the function of mast cells.

Contrary to A 1 and A 2a receptors, where the tissue distribution was detected on the protein level, the presence of A 2b and A 3 receptors was detected on the basis of their mRNA level. Expression levels for A 3 adenosine receptors are rather low comparing to other subtypes and highly species dependent. A 3 adenosine receptors are expressed primarily in the central nervous system, testis, immune system and appear to be involved in the modulation of mediator release from mast cells in immediate hypersensitivity reaction.

The A 3 antagonists published so far in the literature belong to the groups of flavonoides, 1,4-dihydropyridine derivatives, triazoloquinazolines, thiazolonaphthyridines and thiazolopyrimidines. The present invention relates to a novel type of effective A 3 antagonists, which have the aminoquinoline structure.

For therapeutic use it is essential to ensure that the molecule does not bind, or bind only in the case of very high concentration to the A 1 , A 2a and A 2b sub-types of the adenosine receptor. Our present invention relates to the compounds of the general formula (I) as well as their salts, solvates and isomers which have great selectivity for the A 3 subtype of the adenosine receptor.

Our aim was to prepare A 3 ligands first of all with quinoline structure, and within those preferably antagonists, which have strong antagonistic effect and show high selectivity for the A 3 receptor, ie. they inhibit the A 3 receptor in much lower concentration than they inhibit the A 1 , A 2a and A 2b receptors. Further aims were to have stability, bioavailability, therapeutic index and toxicity data which make possible to develope the new compounds into drug substances and that due to their favourable enteral absorbtion the compounds can be applied orally.

We have found that the compounds of the general formula (I)—wherein

R 1 stands for hydrogen atom or a straight or branched C 1-4 alkyl group; R 2 stands for hydrogen atom or a straight or branched C 1-4 alkyl group; R 3 stands for hydrogen atom or a straight or branched C 1-4 alkyl group, or a phenyl group, thienyl group, or furyl group, optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom, or for a 5- or 6 membered heteroaromatic ring—containing one, two or three nitrogen atoms or one nitrogen atom and one oxygen atom or one nitrogen atom and one sulphur atom—  optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom; R 4 and R 5 stand for hydrogen atom or form together an 1,3-butadienyl group, optionally substituted by a methylenedioxy group or one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, hydroxy group or halogen atom; R 6 stands for hydrogen atom or a cyano group, aminocarbonyl group, C 1-4 alkoxycarbonyl group, or carboxy group; R 7 stands for hydrogen atom or a straight or branched C 1-4 alkyl group, or a phenyl group, benzyl group, thienyl group or furyl group, optionally substituted by a methylenedioxy group, or one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, hydroxy group, trifluoromethyl group, cyano group or halogen atom, or for a 5 or 6 membered heteroaromatic ring—containing one, two or three nitrogen atoms or one nitrogen atom and one oxygen atom or one nitrogen atom and one sulphur atom—optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom, X stands for a —CH 2 — group, —NH— group, —NR 8 — group, or a sulphur atom or an oxygen atom or a sulpho group or a sulphoxy group—wherein R 8 stands for a straight or branched C 1-4 alkyl group or C 3-6 cycloalkyl group—; n stands for zero, 1 or 2—and their salts, solvates, and isomers and the salts, solvates of the latter, fulfill the above criteria.

›Detailed meanings of the above listed substituents are…

Detailed meanings of the above listed substituents are as follows:

By a straight or branched C 1-4 alkyl group we mean methyl-, ethyl-, propyl-, isopropyl-, butyl-, isobutyl-, secondary-butyl-, tertiary-butoxy-, preferably ethyl- or methyl group.

By a straight or branched C 1-4 alkoxy group we mean methoxy-, ethoxy-, propoxy-, isopropoxy-, butoxy-, isobutoxy-, secondary-butoxy-, tertiary-butoxy-, preferably ethoxy- or methoxy group.

By a C 3-6 cycloalkyl group we mean cyclopropyl-, cyclobutyl-, cyclopentyl- or cyclohexyl group.

By 1,3-butadienyl-group we mean (—CH═CH—CH═CH—)-group, ie. the pyridine ring substituted by R 4 and R 5 substituents means a benzopyridine ring or by its trivial name a quinoline ring.

The heteroaromatic ring containing one or two or three nitrogen atoms means pyrrol, imidazole, pyrazole, 1,2,3-triazole, 1,2,4-triazole, pyridine, pyrimidine, pyridazine, pyrazine and 1,3,4-triazine ring. The ring is optionally substituted by a C 1-4 alkyl, or alkoxy group or by a halogen atom.

The heteroaromatic ring containing one nitrogen atom and one oxygen or sulphur atom means oxazole, isoxazole, thiazole, isothiazole ring. The ring is optionally substituted by a C 1-4 alkyl, or alkoxy group or by a halogen atom.

Salts of the compounds of the general formula (I) mean salts given with inorganic and organic acids and bases. Preferred salts are those given with pharmaceutically accepted acids as for instance hydrochloric acid, sulphuric acid, ethanesulphonic acid, tartaric acid, succinic acid, fumaric acid, malic acid, citric acid, and bases, as for instance sodium hydroxide, potassium hydroxide, ethanolamine.

Solvates mean solvates given with various solvents, as for instance with water or ethanol.

The compounds of the general formula (I) show geometric and optical isomerism, therefore the invention also relates to mixtures of the geometric isomers, to racemic or optically active geometric isomers, as well as to their salts and solvates.

A favourable group of the compounds of the general formula (I) is formed by the compounds of the general formula (IA), wherein

R 1 stands for hydrogen atom or a straight or branched C 1-4 alkyl group; R 2 stands for hydrogen atom or a straight or branched C 1-4 alkyl group; R 3 stands for hydrogen atom or a straight or branched C 1-4 alkyl group, or a phenyl group, thienyl group, or furyl group, optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom, or for a 5- or 6 membered heteroaromatic ring—containing one, two or three nitrogen atoms or one nitrogen atom and one oxygen atom or one nitrogen atom and one sulphur atom—optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom; R 9 , R 10 , R 11 , and R 12 independently mean hydrogen atom or straight or branched C 1-4 allyl group, or straight or branched C 1-4 alkoxy group, or hydroxy group or halogen atom, or R 9 and R 12 stand for hydrogen atom and R 10 and R 11 form together a methylenedioxy group; R 6 stands for hydrogen atom or a cyano group, aminocarbonyl group, C 1-4 alkoxycarbonyl group, or carboxy group; R 7 stands for hydrogen atom or a straight or branched C 1-4 alkyl group, or a phenyl group, benzyl group, thienyl group or furyl group, optionally substituted by a methylenedioxy group, or one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, hydroxy group, trifluoromethyl group, cyano group or halogen atom, or for a 5 or 6 membered heteroaromatic ring—containing one, two or three nitrogen atoms or one nitrogen atom and one oxygen atom or one nitrogen atom and one sulphur atom—optionally substituted by one or more straight or branched C 1-4 alkyl group, straight or branched C 1-4 alkoxy group, or halogen atom, X stands for a —CH 2 — group, —NH— group, —NR 8 — group, or a sulphur atom or an oxygen atom or a sulpho group or a sulphoxy group—wherein R 8 stands for a straight or branched C 1-4 alkyl group or C 3-6 cycloalkyl group—; n stands for zero, 1 or 2—  and their salts, solvates, optically active isomers and the salts, solvates thereof.

A favourable group of the compounds of the general formula (IA) is formed by the compounds wherein

R 1 stands for hydrogen atom, or methyl group; R 2 stands for hydrogen atom, or methyl group;

R 3 stands for phenyl- or thienyl- or furyl group;

R 9 , R 10 , R 11 , and R 12 mean independently hydrogen atom or straight or branched C 1-4 alkyl group, or straight or branched C 1-4 alkoxy group, or hydroxy group or halogen atom, or R 9 and R 12 stand for hydrogen atom and R 10 and R 11 form together a methylenedioxy group; R 6 stands for hydrogen atom, or cyano group; R 7 stands for 4-methoxyphenyl-, 3-methylphenyl-, 3-methoxyphenyl-, 3-thienyl-, or 3-furyl-group, X stands for —NH— group or for oxygen atom and n stands for 1—  and their salts, solvates, optically active isomers and the salts, solvates thereof

Especially favourable are the following compounds complying with the above criteria:

3-methyl-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide; 4-methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide; 3-methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide; 3,4-methylenedioxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide; N-(4-benzylamino-3-cyanoquinolin-2-yl)thiophene-2-carboxamide; N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)thiophene-3-carboxamide; 4-methoxy-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide; 3,4-methylenedioxy-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)-benzamide; N-(4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)furan-2-carboxamide; N-(4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)thiophene-3-carboxamide,  and their salts, solvates, optically active isomers and the salts, solvates thereof.

According to another of its aspects, the present invention also relates to pharmaceutical compositions containing as active principles the compounds of the general formula (I) or their isomers, salts and solvates, which are preferably oral compositions, but inhalable, parenteral and transdermal formulations are also subjects of the invention. The above pharmaceutical compositions may be solids or liquides, such as tablets, pellets, capsules, patches, solutions, suspensions or emulsions. The solid compositions, first of all tablets and capsules are the preferred pharmaceutical forms.

›The above pharmaceutical compositions are prepared by applying…

The above pharmaceutical compositions are prepared by applying usual pharmaceutical excipients and by using standard methods.

The compounds of the general formula (I) can be used in treating pathologies, in the development of which A 3 receptor plays a role.

The compounds of the present invention having selective activity on the A 3 receptor can be used in the therapeutic and/or preventive treatment of disfunctions of the heart, kidney, respiratory system, central nervous system. They inhibit the protective effect of adenosine in growing tumor cells, prevent mast cell degranulation, inhibit the cytokine production, reduce the intraocular pressure, inhibit the TNFα release, inhibit the migration of eosinophils, neutrophils and other immune cells, inhibit the bronchoconstriction and plasma extravasation.

Based on these effects, adenosine A 3 receptor antagonists of the present invention may be therapeutically useful as antiinflammatory, antiasthmatic, antiischemic, antidepressant, antiarrhytmic, renal protective, antitumor, antiparkinson and cognitive enhancing drugs. They also may be useful in the treatment or prevention of miocardial reperfusion injury, chronic obstructive pulmonary disease (COPD) and adult respiratory distress syndrome (ARDS) including chronic bronchitis, pulmonary emphysema or dyspnea, allergic reactions (e.g. rhinitis, poison ivy induced responses, urticaria, scleroderma, arthritis) other autoimmune diseases, inflammatory bowel disease, Addison's disease, Crohn's disease, psoriasis, rheumatism, hypertension, neurogical function disorders, glaucoma and diabetes (K. N. Klotz, Naunyn-Schmiedberg's Arch. Pharmacol. 362:382,2000; P. G. Baraldi és P. A. Borea, TiPS 21:456, 2000).

The compounds of the present invention may be preferable used for the treatment of diseases such as asthma, COPD and ARDS, glaucoma, tumor, allergic and inflammatory diseases, ischemia, hypoxia, arrythmia and renal diseases.

According to another of its aspects, the present invention relates to the use of the compounds of the general formula (I) in the treatment of the above pathologies. Suggested daily dose is 1-100 mg active ingredient depending on the nature and severeness of the disease and on sex, weight etc. of the patient.

Further subject of the invention is the preparation of the compounds of the general formula (I) and of the intermediates of the general formulae (II) (III) and (IV).

The intermediates of the general formulae (II) (III) and (IV) which are used in the preparation process according to the invention, are partly novel. Substituents of the general formulae (II), (III) and (IV) have the meanings as defined above.

In the process according to our invention the bis-carboxamide of the general formula (II) is selectively hydrolysed and the resulting compound of the general formula (I) is, if desired, transformed into its salts, solvates or, liberated from its salt, solvate and separated into its geometric or optical isomers.

Substituents of the compounds of the general formula (I) may be transformed into each other by known methods.

Selective hydrolysis is performed by using alcoholic, preferably methanolic alkali hydroxide solution, preferably potassium and/or sodium hydroxide solutions, but other agents helping the hydrolysis of amides can also be used.

The selective hydrolysis can be carried out in a wide temperature range, favourably between 20° C.-100° C.

The compounds of the general formula (II)—wherein the meanings of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , X and n are as defined above—can be obtained by several known methods, among them the one demonstrated in Scheme 1 (FIG. 6 ), by acylation of the compounds of the formula (III), by using an acylation method known in the organic chemistry. For acylating agent preferably acyl chloride, for acid binding agent triethylamine and/or pyiridine can be applied, but other acid binding agents can also be used.

The compounds of the general formula (III)—wherein the meanings of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 8 , X and n are as defined above—can be prepared from the compounds of the formula (IV)—by using methods known per se (Nan Zhang, Bioorg. and Med. Chem. Lett., 10,2825, 2000).

The compounds of the general formula (IV)—wherein the meanings of R 4 , R 5 and R 6 are as defined above—can be prepared from the compounds of the formula (V), by using methods known per se (D. L. Leysen, J. Heterocyclic Chem., 24, 1611, 1987).

The compounds of the general formula (V)—wherein the meanings of R 4 , R 5 and R 6 are as defined above—can be prepared from the compounds of the formula (VI), by using methods known per se (Pfizer (Inc) U.S. Pat. No. 4,175,193).

The compounds of the invention, of the general formulae (I), (II), (III) and (IV), their preparation and biological activity are demonstrated in the following Examples, without limiting the scope of claims to the Examples.

FIG. 1 shows general formula (I),

FIG. 2 shows general formula (IA),

FIG. 3 shows general formula (II),

FIG. 4 shows general formula (III) and

FIG. 5 shows general formula (IV).

FIG. 6 shows Scheme 1 the reaction route for the preparation of compounds of the general formula (I).

EXAMPLES
›Examples11
›Example 1

3-Methyl-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide

In general formula (I) R 1 and R 2 stand for hydrogen atoms, R 3 for phenyl group, R 4 and R 5 form together a 1,3-butadienyl group, R 6 stands for cyano group, R 7 for 3-methylphenyl group, the meaning of X is —NH group, n is 1.

a.) 2-Amino-3-cyano-4-chloroquinoline

The mixture of 10 g of 2-amino-3-cyano-4-hydroxyquinoline and 15 ml of phosphoryl chloride is heated under stirring at 110 ° C. The reaction mixture is cooled down, poured onto 100 ml of ice-water and neutralized with 60 ml of 10% sodium hydroxide solution. The resulting yellow precipitate is filtered off, washed with 50 ml of water. After drying 7.5 g of the title compound is obtained, mp.: 210° C.

NMR, δ H (400 MHz, DMSO-d 6 ): 7.21 ppm, (s, 2H, NH 2 ), 7.35-7.40 ppm, (dd, 1H, 6-H), 7.53-7.57 ppm, (d, 1H, 5-H), 7.70-7.75 ppm, (dd, 1H, 7-H), 7.93-7.98 ppm, (d, 1H, 8-H)

b.) 2-Amino-3-cyano-4-benzylaminoquinoline

5 g of 2-amino-3-cyano4-chloroquinoline and 11 ml of benzylamine are heated under stirring at 130° C. The reaction mixture is poured onto 50 ml of water, the resulting precipitate is filtered off, washed with 50 ml of water. The pale-yellow precipitate is recrystallized from dimethylformamide to obtain 5.2 g of the title compound.

Mp.: 206° C.

NMR, δ H (400 MHz, DMSO-d 6 ): 5.02-5.03 ppm, (d, 2H, N—CH 2 ), 6.22 ppm, (s, 2H, NH 2 ), 7.14-7.16 ppm, (dd, 1H, 6-H), 7.24-7.26 ppm, (dd, 1H, 5-H), 7.30 ppm, (s, 5H, Ph), 7.50-7.52 ppm, (dd, 1H, 7-H), 8.16-8.19 ppm, (d, 1H, 8-H), 8.30-8.33 ppm, (t, 1H, NH)

Using 2-aminomethylpyridine or 3-aminomethylpyridine or 4-aminomethylpyridine instead of benzylamine, the appropriate compounds of general formula III can be obtained.

c.) 3-Methyl-N-(3-methylbenzoyl)-N-(4-benzylamino-3-cyanoquinoline-2-yl)benzamide

To the solution of 5 g of 2-amino-3-cyano-4-benzylaminoquinoline in 30 ml of pyridine 6 ml of 3-methylbenzoyl chloride are dropped, under stirring at 0° C. The reaction mixture is stirred at 80° C. for 8 hour, then it is poured onto 150 ml of ice-water. The precipitate is filtered off, washed twice with 40 ml of water. The resulting white crystalline material is recrystallized from 200 ml of ethanol to give 9.2 g of the title compound, mp.: 234° C.

By using pyridine-3-carbonyl chloride as acylating agent, the appropriate compound of general formula II can be obtained.

d.) 3-Methyl-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide

To the solution of 5 g of 3-methyl-N-(3-methylbenzoyl)-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide in 80 ml of acetonitrile 20 ml of 1N methanolic potassium hydroxide solution are added. The reaction mixture is refluxed for 3 minutes, then 3 ml of glacial acetic acid is added to it, then it is neutralized with 50 ml of 1M sodium hydrogen carbonate solution and the resulting crystals are filtered off. The white crystalline material is recrystallized from 130 ml of acetonitrile to give 3.1 g of the title compound of general formula (I). Mp.: 230° C.

›Example 2

4-Methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is phenyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 4-methoxyphenyl group, X means —NH— group, n is 1.

2-amino-3-cyano-4-benzylaminoquinoline, prepared as described in Example 1, is transformed with 4-methoxybenzoyl chloride, analogously as described in Example 1, into 4-methoxy-N-(4-methoxybenzoyl)-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide, which after selective hydrolysis, by the method described in Example 1, results the title compound of general formula (I). Melting point of the title compound: 188° C.

Sodium salt of the title compound is prepared by the following method:

4-methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide is dissolved in methanol and equivalent amount of sodium hydroxide in methanol is added to it. The precipitated white crystalline material is filtered off. Mp.: 255° C.

Ethanesulfonate salt of the title compound is prepared by the following method: 4-methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide is dissolved in methanol and equivalent amount of ethanesulfonic acid is added to it. The precipitated white crystalline material is filtered off. Mp.: 223° C.

›Example 3

3-Methoxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is phenyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 3-methoxyphenyl group, X means —NH— group, n is 1.

2-amino-3-cyano-4-benzylaminoquinoline, prepared as described in Example 1, is transformed with 3-methoxybenzoyl chloride, analogously as described in Example 1, into 3-methoxy-N-(3 -methoxybenzoyl)-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide, which after selective hydrolysis by the method described in Example 1, results the title compound of general formula (I). Melting point of the title compound: 186° C.

›Example 4

3,4-Methylenedioxy-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is phenyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R7 means 3,4-methylenedioxyphenyl group, X means —NH— group, n is 1.

2-amino-3-cyano-4-benzylaminoquinoline prepared as described in Example 1, is transformed with 4-methoxybenzoyl chloride analogously as described in Example 1, into 3,4-methylenedioxy-N-(3,4-methylenedioxybenzoyl)-N-(4-benzylamino-3-cyanoquinolin-2-yl)benzamide which after selective hydrolysis by the method described in Example 1, results the title compound of general formula (I).

Melting point of the title compound: 231° C.

›Example 5

N-(4-benzylamino-3-cyanoquinolin-2-yl)thiophene-2-carboxamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is phenyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 2-thienyl group, X means —NH— group, n is 1.

2-amino-3-cyano-4-benzylaminoquinoline prepared as described in Example 1. is transformed with thiophene-2-carbonyl chloride, analogously as described in Example 1, into N-(2-thiophenecarbonyl)-N-(4-benzylamino-3-cyanoquinolin-2-yl)thiophene-2-carboxamide, which after selective hydrolysis, by the method described in Example 1, results the title compound of general formula (I).

Melting point of the title compound: 197° C.

›Example 6

N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)thiophene-3-carboxamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is 2-thienyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 3-thienyl group, X means —NH— group, n is 1.

a) 2-amino-3-cyano4-(2-thienylmethylamino)quinoline

5 g of 2-amino-3-cyano4-chloroquinoline, prepared as described in Example 1, is stirred with 11 ml of 2-thienylmethylamine at 130° C. for 3 hours. The reaction mixture is poured onto 50 ml of water, the resulting precipitate is filtered off, washed with 50 ml of water. The pale yellow material is recrystallized from 25 ml of ethanol to obtain 5.2 g of title compound, mp.: 208° C.

The 2-amino-3-cyano-4-(2-thienylmethylamino)quinoline prepared as described above is transformed with thiophene-3-carbonyl chloride, analogously as described in Example 1, into N-(3-thiophenecarbonyl)-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)-thiophene-3-carboxamide which after selective hydrolysis, by the method described in Example 1, gives the title compound of general formula (I). Melting point of the title compound: 223° C.

›Example 7

4-methoxy-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is 2-thienyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 4-methoxyphenyl group, X means —NH— group, n is 1.

The 2-amino-3-cyano-4-(2-thienylmethylamino)quinoline prepared as described in Example 6. is transformed with 4-methoxybenzoyl chloride into 4-methoxy-N-(4-methoxybenzoyl)-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide by the method described in Example 1, which after selective hydrolysis gives the title compound of general formula (I). Melting point of the title compound: 173° C.

›Example 8

3,4-methylenedioxy-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)-benzamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is 2-thienyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 3,4-methylenedioxyphenyl group, X means —NH— group, n is 1.

2-amino-3-cyano-4-(2-thienylmethylamino)quinoline prepared as described in Example 6. is transformed with 3,4-methylenedioxybenzoyl chloride into 3,4-methylenedioxy-N-(3,4-methylenedioxybenzoyl)-N-(4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide by the method described in Example 1, which after selective hydrolysis gives the title compound of general formula (I). Melting point of the title compound: 241° C.

›Example 9

N-(4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)furan-2-carboxamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is 2-furyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 2-furyl group, X means —NH— group, n is 1.

a.) 2-Amino-3-cyano4-(2-furylmethylamino)quinoline

5 g of 2-amino-3-cyano-4-chloroquinoline, prepared as described in Example 1 are stirred with 1 ml of 2-furylmethylamine (furfurylamine) at 130° C. for 3 hours. The reaction mixture is poured onto 50 ml of water, the resulting precipitate is filtered off, washed with 50 ml of water. The pale yellow material is recrystallized from 20 ml of ethanol to obtain 4.8 g of the title compound, mp.: 208° C.

The 2-amino-3-cyano-4-(2-furylmethylamino)quinoline prepared as described above is transformed with furan-2-carbonyl chloride by the method described in Example 1. into N-(2-furancarbonyl)-N-(4-[2-furylmethylamino]-3-cyanoquinolin-2-yl) furan-2-carboxamide which after selective hydrolysis gives the title compound of general formula (I). Melting point of the title compound: 196° C.

›Example 10

N-(4-[2-furylmethylamino]-3-cyanoquinolin-2yl)thiophene-3-carboxamide

In the general formula (I) the meaning of R 1 and R 2 is hydrogen atom, R 3 is 2-furyl group, R 4 and R 5 mean together a 1,3-butadienyl group, R 6 means cyano group, R 7 means 3-thienyl group, X means —NH— group, n is 1. 1.

The 2-amino-3-cyano4-(2-furylmethylamino)quinoline prepared analogously as described in Example 6. is transformed with thiophene-3-carbonyl chloride by the method described in Example 1. into N-(3-thiophene carbonyl)-N-(4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)thiophene-3-carboxamide which after selective hydrolysis, performed analogously as described in Example 1. gives the title compound of general formula (I). Melting point of the title compound: 118° C.

Structure and physical characteristics of further compounds of general formula (I) prepared by the method described in Example 1. are shown in Tables I. and II.

Structure and physical characteristics of intermediates of general formula (II) prepared by the method described in Example 1. are shown in Table III.

Structure and physical characteristics of intermediates of general formula (III) and (IIIa) prepared by the method described in Example 1. are shown in Table IV.

Structure and physical characteristics of intermediates of general formula (V) prepared by the method described in Example 1. are shown in Table V.

›Example 172

Tablets of the following composition are made by known methods used in the pharmaceutical industry

Biology

Methods

Human Adenosine A 3 Receptor Binding

Preparing membrane suspension: collect CHO cells expressing hA 3 receptors by washing three times with ice cold PBS, centrifugate at 1000×g 10 min, homogenize for 15 sec in buffer (50 mM Tris, 10 mM MgCl 2 , 1 mM EDTA, pH 8.0), centrifugate at 43,000×g for 10 min (Sigma 3K30), suspense the membrane preparation in the buffer mentioned above, store the aliquots at −80 C.

Binding protocol: incubate CHO-hA 3 membrane preparation (2 μg protein content) in incubation buffer (50 mM Tris, 10 mM MgCl 2 , 1 mM EDTA, 3 U/mL adenosine deaminase, pH 8.0), in the presence of 0.5 nM [ 125 ]AB-MECA (p-amino-benzyl-methylcarboxamido-adenosine) (100.000 cpm) and 100 μM R—PIA (N 6 -[L-2-phenylisopropyl]adenosine) to define non-specific binding or test compound in a total volume of 50 μL for 1 hr at room temperature. Filter over Whatman GF/B glass fibre filters (presoaked in 0.5% polyethylenimine for 3 hours), wash 4× with 1 mL ice-cold 50 mM Tris, 10 mM MgCl 2 , 1 mM EDTA (pH 8.0) on 96-well Brandel Cell Harvester. Detection of activity: in gamma-counter (1470 Wizard, Wallac). Inhibition [%]=100−((activity in the presence of test compound−non-specific activity)/(total activity−non-specific activity))*100

Human Adenosine A 1 Receptor Binding

Preparing membrane suspension: collect CHO cells expressing hA 1 receptors by washing three times with ice cold PBS, centrifugate at 1000×g 10 min, homogenize for 15 sec in buffer (50 mM Tris, pH 7.4), centrifugate at 43.000×g for 10 min (Sigma 3K30), suspense the membrane preparation in the buffer mentioned above, store the aliquots at −80° C.

Binding protocol: incubate CHO—hA 1 membrane preparation (50 μg protein content) in incubation buffer (50 mM Tris, 3 U/mL adenosine deaminase, pH 7.4), 10 nM [ 3 H]CCPA (2-chloro-N 6 -cyclopenthyl-adenosine) (80,000 dpm) and 10 μM R-PIA (N 6 -[L-2-phenylisopropyl]adenosine) to define the non-specific binding or test compound in a total volume of 100 μL for 3 hr at room temperature. Filter over Whatman GF/B glass fibre filters (presoaked in 0.5% polyethylenimine for 3 hours), wash 4× with 1 mL ice-cold 50 mM Tris (pH 7.4) on 96-well Brandel Cell Harvester. Detection of activity: in 96-well plate in the presence of HiSafe-3 cocktail in beta-counter (1450 Microbeta, Wallac). Inhibition [%]=100−((activity in the presence of test compound−non-specific activity)/(total activity−non-specific activity))*100

Human Adenosine A 2a Receptor Binding

Binding protocol: incubate 7 μg of membranes (human A 2a adenosine receptors transfected into HEK-293 cells, source: Receptor Biology, Inc.), buffer (50 mM Tris-HCl, 10 mM MgCl 2 , 1 mM EDTA, 2 U/mL adenosine deaminase, pH 7.4), 20 nM [ 3 H]CGS-21680 (2-[p-(2-carbonylethyl)phenylethylamino]-5′-N-ethylcarboxamido-adenosine) (200.000 dpm) and 50 μM NECA (5′-N-ethylcarboxamido-adenosine) to define the non-specific binding or test compound in a total volume of 100 μl for 90 min at room temperature. Filter over Whatman GF/B glass fibre filters (presoaked in 0.5% polyethylenimine), wash 4× with 1 mL ice-cold 50 mM Tris, 10 mM MgCl 2 , 1 mM EDTA, 0.9% NaCl, pH 7.4) on 96-well Brandel Cell Harvester. Detection of activity: in 96-well plate in the presence of HiSafe-3 cocktail in beta-counter (1450 Microbeta, Wallac). Inhibition [%]=100−((activity in the presence of test compound−non-specific activity)/(total activity−non-specific activity))*100

Human Adenosine A 2b Receptor Binding

Binding protocol: incubate 20.8 μg of membranes (human A 2b adenosine receptors transfected into HEK-293 cells, source: Receptor Biology, Inc.), buffer (50 mM Tris-HCl, 10 mM MgCl 2 , 1 mM EDTA, 0.1 mM benzamidine, 2 U/mL adenosine deaminase, pH 6.5), 32.4 nM [ 3 H]DPCPX (8-cyclopenthyl-1,3-dipropylxanthine) (800.000 dpm) and 100 μM NECA (5′-N-ethylcarboxamido-adenosine) to define non-specific binding or test compound in a total volume of 100 μL for 30 min at room temperature. Filter over Whatman GF/C glass fibre filters (presoaked in 0.5% polyethylenimine), wash 4× with 1 mL ice-50 mM Tris-HCl (pH 6.5) on 96-well Brandel Cell Harvester. Detection of activity: in 96-well plate in the presence of HiSafe-3 cocktail in beta-counter (1450 Microbeta, Wallac). Inhibition [%]=100−((activity in the presence of test compound−non-specific activity)/(total activity−non-specific activity))*100

Results

We consider the compounds as biologically active ones if they inhibit the binding of the radioligand on human adenosine A 3 receptors with an activity above 80 % at 1 μM in our experimental conditions.

The dissociation constant (K d ) of [ 125 I]AB-MECA on CHO-hA 3 membrane preparation is determined by isotope saturation studies with the help of Scatchard analysis (G. Scatchard, Ann. N.Y. Acad. Sci. 51:660, 1949). The IC 50 is converted to an affinity constant (K i ) by application of the Cheng-Prusoff equation (Y. J. Cheng and W. H. Prusoff, Biochem. Pharmacol. 22:3099, 1973).

Several compounds of the general formula (I), (II), (III) and (IV) display remarkable biological effects. The compounds of the general formula (IA), defined in claim 2 , as a subgroup of the general formula (I), defined in claim 1 , exert the most important activities. Except of 5 compounds, their K i values are not higher than 20 nM. The compounds given as examples are especially advantageous. Their K i values in human adenosine A 3 receptor binding studies are between 0.19 and 0.69 nM. The K i values of the most advantageous compounds are 0.14 and 0.15 nM.

The compounds possess proper bioviabilities and exert at least 10,000-fold selectivity in respect of human adenosine A 1 , A 2a and A 2b receptor subtypes.

Further, the duration of their action at intravenous and oral administration is long enough, their ED 50 values are low, their toxicological and side-effect profiles are advantageous.

Data above make the compounds of the general formula (I) probable for therapeutic applications.

›Tables in the description — 3
TABLE I — Mp
No.:XR 3R 6R 7n[° C.]
11.NH
CN
1237
12.NH
CN
1128
13.NH
CN
1116
14.NH
CN
1100,5
15.NH
CN
1223
16.NH
CN
1193,5
17.NH
CN
1193
18.NH
CN
1208
19.NH
CN
1215
20.NH
CN
1250
21.NH
CN
1205
22.NH
CN
1238
23.NH
CN
1212
24.NH
CN
1215
25.NH
CN
1234
26.NH
CN
1160,5
27.NH
CN—Me1184
28.NH
CN
1141,5
29.NH
CN
1194
30.NH
CN
1203
31.NH
CN
1152
32.NH
CN
1190
33.NH
CN
1202
34.NH
CN
1207
35.NH
CN
1159
36.NH
CN
1200
37.NH
CN
1206
38.NH
CN
1221
39.NH
CN
1198
40NH
CN
1158
41.NH
CN
1178
42.NH
CN
1198,5
43.NH
CN
1197,5
44.NH
CN
1191
45.NH
CN
1168,5
46.N-Me
CN
1155
47.NH
H
1172
48.NH
H
1250
49.NH
H
1264
50.NH
H
1265
51.NH
H
1163
52.O
CN
1157
53.S
CN
1196
54.S═O
CN
1205
55.NHHCN
0266
56.NH
CN
1154
57NH
CN
1145
TABLE IV — Mp
No.:R 1R 2R 3R 4R 5Xn[° C.]
129HH
NH1192
130HH
NH1202
131HH
NH1250
132HH
NH1167
133H
NH1183
134H
NH1182
135HH
NH2172
136HH
NH2143
137H
NH2129
138H
NH2136
139HH
N—Me1212
140HH
S1168
141HH
O1213
142HH
NH1234
143HH
NH1221
144HH
NH1198
145HH
NH1201
146HH
NH1213
147HH
NH1198
147HH
NH1201
148HH
NH1167
149HH
NH1156
150HH
NH1187
151HH
NH1178
152HH
NH1207
153HH
NH1217
154HH
NH1204
155HH
NH1216
156HH
NH1205
158HH
NH1213
159HH
NH1200
160
NH0214
Active ingredient25 mg
Lactose50 mg
Avicel21 mg
Crospovidone3 mg
Magnesium stearate1 mg
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39 codes
IPC · International Patent Classification
Section A — Human necessities
  • A61K/
  • A61P11/00
  • A61P13/12
  • A61P27/06
  • A61P43/00
  • A61P17/06
  • A61P11/06
  • A61P/
  • A61K31/4706
  • A61P25/16
  • A61P9/12
  • A61K31/4709
  • A61P25/00
  • A61P37/08
  • A61P25/24
  • A61P9/00
  • A61P29/00
  • A61P3/10
  • A61P35/00
  • A61P9/10
Section C — Chemistry; metallurgy
  • C07D213/85
  • C07D409/14
  • C07D215/48
  • C07D405/14
  • C07D409/12
  • C07D/
  • C07D405/12
  • C07D213/75
  • C07D401/12
  • C07D215/46
  • C07D215/54
USPC · US Patent Classification
514/313546/159546/161514/352546/308514/353546/279.7546/160

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